Hydrazo steroids and method of manufacture



Patented Apr. 15, 1969 jugated diene. A preferred species of this composition 3,438,974 aspect are the 1,4-adduct of an azodicarboxylic acid lower HYDRAZO g gg gg ggg F alkyl ester or amide with a 16,20-pregnadiene. Munemitsu Tomoeda, Kanazawa, Japan, assignm. to The invention sought to be patented In another of its Schering Corporation Bl fi ld NJ a col-pm composition aspects resldes 1n the concept of an allylic ti f N Jersey adduct of an azodicarboxylic lower alkyl ester with a N0 Drawing. Continuation-impart of application Ser. No. steroid homo cisoid conjugation diene.

1965- This application May 6, 1966, Virtually any steroid of the androstane, pregnaue, or 543,035 cholestane series, for example, which possesses a homo Int. Cl. C07c 173/10, 167/06 U S Cl 5 17 Claims cisoid conjugated diene system will serve as the starting material for eifecting the process aspect of this invention to produce a tangible embodiment of the product aspects. By homo cisoid is meant that the double bonds ABSTRACT OF THE DISCLOSURE are conjugated between carbon atoms in the cis spatial The process of reacting an azodicarboxylic acid ester configuration. For the most part, a homo cisoid conor an equivalent thereof with a steroid homo cisoid conjugated diene will be homo annular, that is, the conjugated diene in a non-reactive solvent yields a hydrazo jugated double bonds of the starting material will be consteroid of the group consisting of a 1,4-adduct and an altained within the same ring of the condensed 4-ring steroid lylic adduct of said steroid homo cisoid conjugated diene nucleus such as exemplified by a 1,3-diene, a 2,4-diene, a

with said azodicarboxylic acid ester or equivalent thereof, -d nc, a ,3(9)-diene, a 8(14), 9(11)-diene, a 8(9), which hydrazo steroids have bactericidal, bacteriostatic 11(l2)-diene, a 14,16-diene and the like. In certain inand fungicidal properties. Also useful as anti-androgens stances, the conjugated diene system satisfies the requireare a preferred species of hydrazo steroids, i.e. 20-lower ments of being homo cisoid when it is not homo annular alkanoyloxy 16,21 hydrazo 17(20) pregnene N such as is evidenced by the l6,2()-diene system or the 7, N -dicarboxylic acid lower alkyl esters (and amides) and 14-dieue system.

the corresponding 20-hydroxy derivatives thereof, said 20- The following scheme depicts the general reaction of hydroxy derivatives usually being isolated in the tauthe process aspect of the invention.

COOCHB C2H5OOCN CzH5OOC-N CO0C2H C2H5O0C-NH I II III IV tomeric ZO-keto form as 20-keto-16,21-hydrazo-17 (a and In the foregoing scheme, only the four-ring nucleus of the B)-pregnane-N ,N -dicarboxylic acid lower alkyl esters starting steroid homo cisoid conjugated diene is shown for (or amides). the sake of simplicity. At C-1O and/0r C-13, hydrogen may be present or an angular substituent such as a lower This application is a continuation-in-part of copending alkyl group and preferably methyl substiments as application sen NO. 438 100 filed Man 8 1965 now abam scribed later herein may appear on the nucleus and side doned, of Munemitsu Tomoeda for Hydrazo Steroids and cham at the 9 q A i 1S depleted solely Method of Manufacnna by way of exemplificatlon for it is apparent that a homo cisoid conjugated diene may be present in a finite number of other places in the steroid molecule. In the 10,13- dimethyl steroid group of compounds represented by androstanes, pregnanes, cholestanes, coprostanes, and the like, the diene system may be present at the following positions: (1,3), (2,4), (5,7), (6,8), (7,14), (8,11), (8(14)), 9(11)), (14,16), and (16,20). In the 19-nor This invention relates to compositions of matter classified in the art of chemistry as hydrazo steroids and to methods for the manufacture of and use of such compositions.

The invention sought to be patented in its process aspect is defined as residing in the concept of preparing a hydrazo steroid of the group consisting of 1,4- and allylic f h 1 adducts of a steroid homo cisoid conjugated diene with analogs of the Oregomg t e lane System may a so be an azodicarboxylic lower alkyl ester, or a hereinafter dis- Present at the followmg Posltlons: (1(10)2) (1(1O)4) closed equivalent thereof, by admixing a steroid homo 901)) (15(10)) (35(1O)) (5(10)6) (53) cisoid conjugated diene with the aforementioned dieno- (79) (5(10,)8); and m the lis'nor analogs of the phile in an inert organic solvent at a temperature in the foregomg the (heme Systems may also be Present at range of from about room temperature to the reflux tem- X Analogous fqrmatiofl f a d ct or allperature of th ti i yl1c adduct occurs concomitant wlth analogous shifting of The invention sought to be patented in one of its com- 69 the double bondsposition aspects resides in the concept of the 1,4-adduct Stated he et fore, any Steroid possessing the basic of an azodicarboxylic lower alkyl ester, or its hereinafter r q n f a h m CI Old Conjuga ed diene IS 3. Suitdisclosed equivalent, with a steroid homo cisoid conable starting material for my reaction. The steroid may bear substituents without limit provided such substituents do not react with the dienophile, II. Carbonyl groups if present and reactive to the dienophile are preferably first protected as ketals or acetals While hydroxy or amino groups are preferably first blocked by means of an acylating agent such as acetic or benzoic anhydride or chloride forming an ecyl ester or amide, while carboxyl groups are preferably esterified. Thus carbonyl groups, preferably protected by ketalization (or acetal formation) may be present at positions such as C-3, '11, 17, 20, 21, for example, hydroxyl groups preferably in the form of lower alkanoyl esters may be present at positions C-3, 11, 12, l4, 16, 17, 20, 21, for example. Similarly, amino and carboxy groups, also preferably protected as described above, may appear at various places in the steroid. Groups such as alkyl containing up to 4 carbons and preferably methyl, may apear at positions Cl, 2, 6, 7, 10, ll, 13, 16, 17 and 21. Indeed, even longer chains may appear at C-17 as evidenced by the cholestane series. The alkyl groups do not have an effect upon the reaction. Similarly, halogeno groups, such as fluoro, chloro, bromo and iodo may be present at various positions, for example, at C-6 and C-9, without effecting the course of the reaction. In addition, ester and ether groups may be present since they have little or no effect on the course of the reaction.

The starting materials, or their derivatives, are known in the art or may be prepared by methods established in the art. Representative of such starting materials are:

2,4-cholestadiene,

5,7-androstadiene-3fi,17fl-diol 3,17-diacetate,

-5,7-androstadiene-3;3,17B-diol 3-acetate 17-benzoate,

5,7-androstadiene-3fl,17,3-diol 3,17-dibenzoate,

3 B-hydroxy-5,7-androstadien-17-one 3-acetate,

3 fi-hydroxy-S,7-androstadien-17-one 3-benzoate,

3B-hydroxy-5,7-pregnadien-20-one B-acetate,

3B-hydroXy-5,7-pregnadien-20-one 3-benzoate, 313,21-dihydroxy-5,7-pregnadien-2O-one 3,2l-diacetate, 3,8,21-dihydroxy-5,7-pregnadien-20-one 3,2 l-dibenzoate, 5,7-norcholestadien-3fi-ol 3-acetate, 5,7-norcholestadien-35-01 3-benzoate, 5,7-cholestadiene, 7-dehydrocholesterol 3-acetate,

7-dehydrocholesterol 3-methyl ether,

7-dehydrocholesterol 3-benzoate,

22-iso-5,7-spirostadien-3 3-01 3-acetate, 22-iso-5,7-spirostadien-313-01 3-benzoate,

3fi-acetoxy methyl-5,7-cholestadiene,

methyl 5 ,7 -cholestadiene-3-carb0xylate,

ergosterol 3-acetate, ergosterol 3-benzoate,

6,8-coprostadien-3fi-ol 3-acetate,

methyl 3a,125-dihydroxy-7,l4-choladienoate 3,l2-

diacetate, cholesterol-B 3-acetate,

ergosterol-B 3-acetate,

3 a-acetoxy- 1 6,20-pregnadien-1 l-one,

3,6-acetoxy-l6-allopregnen-20-one 20-enol acetate (i.e.

3,8-3C6IOXY-5OL-1 6-pregnen-20-one 20-enol acetate),

SB-acetoxy-S, 1 6-pregnadien-20-one 20-enol acetate,

3 ,8-benzyloxy-5,16-pragnadien-20-one 20-enol acetate,

5 ,l6,20-furostatriene-3 19,26-diol 3,26-diacetate,

l6-anhydrogitoxigenin acetate,

fi-(3 ,B-acetoxy-l6-etioallocholenyl-17)-A' -butenolide,

3a-acetoxy-16-pregnen-20-one -enol acetate,

3,6-acetoxy-16-allopregnene-l1,20-dione 20-enol acetate (i.e. 3fi-acetoxy-5e-16-pregnene-11,20-dione 20-enol acetate),

3 ,B-acetoxy-l6-allopregnene-l 1,20-dione 9 11 ),20-bisenol acetate (i.e. 3 fi-acetoxy-S a-16-pregnene-1 1,20-dione 9(1 1),20-bisenol acetate),

313,1lm-diacetoxy-l6-allopregnen-20-one 20-enol acetate (i.e. 36,1 la.-acetoxy-5a-16-pregnen-20-one 20-eno1 acetate),

3a-acetoxy-l6-pregnen-1l,20-dione 20-enol acetate,

3a,1lfi-diacetoxy-l6-allopregnen-20-one 20-enol acetate (i.e. ,1l,S-diaCetoXy-Sa-16-pregnen-20-one 20-enol acetate).

Included within the foregoing are the ketals, e.g. ethylene ketal, of the carbonyl containing steroids, said ketals being preparable by standard methods well known in the art.

In addition to the foregoing, A -steroids of the androstane and pregnane series as described for example in U.S. Patents 3,082,220, 3,086,012, 3,086,013, and 3,138,619 as well as A -dehydro androstanes as described in U.S. Patent 3,096,353 may serve as starting materials with or without further derivatization (protection of functional groups).

The dienophilic azodicarboxylic ester, II, is the preferred reactant for reasons of its ease of availability and preparation. Other lower alkyl esters such as methyl and propyl are considered full equivalents. Also considered as an equivalent of the aforementioned dienophiles are azodicarboxylic amides and their N-lower alkyl analogs. These behave in analogous fashion to the aforementioned azodicarboxylic esters giving rise to adducts possessing an amide function in place of the carbalkoxy group. The reaction is carried out in a non-reactive solvent which will dissolve the reactants and possess a boiling point in the range of about 50 to C. Preferably the solvent is benzene and the temperature of the reaction established by refluxing the mixture until substantial reaction has occurred, usually less than 30 hours. Other solvents which may be employed are hydrocarbons such as toluene, heptane; ethers such as tetrahydrofuran, dioxane, di-isopropyl ether, di-n-butyl ether; esters such as ethyl acetate, methyl propionate and the like; nitriles such as acetonitrile, propionitrile; amides such as dimethylformamide, dimethylacetamide and the like.

The product(s) of the reaction are removed by usual techniques such as fractional crystallization or chromatographic separation. The products of the reaction, namely the 1,4-adduct as exemplified by III, supra, and the allylic adduct as exemplified by IV, supra, are formed in varying ratios to each other depending upon the spatial characteristics and substitution characteristics of the starting diene.

The products of the reaction, namely the 1,4-adduct as represented by III and the allylic adduct, as represented by IV, have valuable bactericidal, bacteriostatic and fungicidal properties especialy against gram positive organisms. They are thus useful as germicides and fungicides. Compositions comprising such products may be used in solutions, suspensions of solid mixtures to sterilize laboratory equipment, hospital areas and the like. In addition, the products of the reaction are useful intermediates and building blocks for preparing other pharmacologically active substances. For example, the products may be quaternized with lower alkyl halides or sulfates thus forming quaternary salts which themselves are germicidal substances. Further various chemical transformations such as reduction of the ester groups to hydroxyl or removal thereof gives rise to steroidal amines which lend themselves to additional transformations.

In addition to the foregoing, useful as anti-androgens are a preferred species of one of the composition aspects of my invention, namely 20-lower alkanoyloxy-16,21-hydrazo 17(20) pregnene-N ,N -dicarboxylic acid lower alkyl esters (and amides) (Formula V below wherein R is lower alkanoyloxy) as well as the corresponding 20-hydroxy derivatives thereof (Formula V wherein R is hydrogen). The 20-hydroxy derivatives are isolated in the more stable tautomeric 20-keto form (Formula VI below) as 20 keto 16,21-hydrazo-17(u and ,B)-pregnane-N ,N -di carboxylic acid lower alkyl esters (or amides).

The 16,21 hydrazopregnane N ,N -dicarboxylic acid derivatives of my invention, useful as anti-androgens, include derivatives defined by following Formulae V below and the S-dehydro analogs thereof, it being understood that when R is hydrogen, the 16,2l-hydrazo-N ,N -dicarboxylic acid pregnane derivatives (V') are isolated in the more stable tautomeric form as ZO-ketO-17(oc and 5)- pregnanes as shown in Formula VI below.

LOO, mu ii wherein R is an acid radical of a hydrocarbon carboxylic acid having up to 8 carbon atoms; Y is a member selected from the group consisting of hydrogen, keto, (H,aOR) and (H,fiOR); R and R" are members selected from the group consisting of hydrogen and lower al-kanoyl; and Z is a member selected from the group consisting of amido and lower alkoxy.

By lower alkyl are contemplated hydrocarbon radicals having up to four carbon atoms, preferably methyl and ethyl.

By lower alkoxy are thus contemplated alkoxy radicals having up to four carbon atoms.

Included within the term acid radical of a hydrocarbon carboxylic acid having up to 8 carbon atoms are acid radicals of lower alkanoic acids including propionic, butyric, valeric, caproic, and, preferably, acetic; as well as aryl carboxylic acids such as toluic acid and, preferably, benzoic acid.

In the above formulae, the use of a wavy line indicates that the group attached by a bond so drawn may exist in either an alpha or beta position relative to the nuclear carbon to which it is joined. Thus, a 20-enolacetate-16,2l-hydrazo derivative of my invention (V) exists in two isomeric forms, namely, as 16a,21-hydrazo and 165,21-hydrazo derivatives. Throughout this specification and in the claims, when a configuration is not specified in a compound name, it is understood that both isomeric forms are included. Thus the compound name, diethyl 3B benzoyloxy 20 acetoxy 16,21 hydrazo- 5,17(20) pregnadiene-N ,N -dicarboxylate- (a compound of Formula V wherein R is benzoyl, R is acetyl, Y is hydrogen, and Z is OC H implicitly includes both the 1611- and 165-isomers, namely, diethyl 3 3-benzoyloXy-20- acetoXy 16m,21-hydrazo-5,17(20)-pregnadiene-N ,N -dicarboxylate and diethyl 318-12enzoyloxy-ZO-acetoxy-1618,21- hydrazo-5,17 (20)-pregnadiene-N ,N -dicarboxylate. Similarly, the compound name, diethyl 3/3-benzoy1oxy-20-keto- 16,21 hydrazo 17(a and B)-5-pregnene-N ,N dicarboxylate (a compound of Formula VI wherein R is benzoyl, Y is hydrogen, and Z is OC H inherently includes the 16a,17aand 165,17B-isomeric forms, namely diethyl 3/3-benzoyloxy keto-16u,21-hydrazo-17a-5- pregnene-N ,N -dicarboxylate and diethyl 3fi-benzoyloxy- ZO-keto-I65,21-hydrazo-5-pregnene N N -dicarboxylate (it being understood when no configuration is given for the C-17 side chain that it is in the beta position).

Additionally, throughout this specification and in the claims when a name for a pregnane saturated at C-5 does not indicate a specific configuration, it is understood that the hydrogen at C-5 is in the normal, i.e. S-B-position. Thus, the name 3a-acetoxy-16-pregnen-20-one 20- enol acetate indicates that the hydrogen at C-5 is in the beta position, and the compound might also be named 3a-acetOXy-Sfl-16-pregnen-20-one 20-enol acetate.

Included in the preferred group of compounds (V) are the 1,4-adducts of azodicarboxylic acid ethyl ester with ZO-acetoxy-16,20-pregnadienes such as:

(1') Diethyl 3,9,20diacetoxy-16,21-hydrazo-5,17(20)- pregnadiene-N ,N dicarboxy1ate,

(2) 3B benzoyloxy-S,16-pregnadien 20-one ZO-enol acetate,

(3) 3a-acetuxyJfi-l6-pregnene-11,20-dione 20 enol acetate,

(4) 3 3 acetoxy 5a-16-pregnene11,20-dione 20-enol acetate,

(5) 3a-acetoxy-5,B16-pregnen-20-one 20-enol acetate,

(6) 3fi-acetoXy-5u-16-pregnen-20-one 20-enol acetate,

(7) 35,11 diacetoXy-5a-9(1l),l6-pregnadien 20-0ne 20-enol acetate,

(8) 35,110; diacetoxy-5a-16-pregnen 20-one 20-enol acetate, and

(9) :,115 diacetoxy -16-pregnen-20-0ne ZO-enol acetate, each of which adducts are derived in the 20-enol acetate form (i.e. Formula V wherein R is acetyl, Z being ethoxy) and are named, respectively, as follows:

(1') Diethyl 3B,20'diacetoxy-16,21-hydrazo 5,17(20) pregnadiene-N ,N -dicarboxylate,

(2) Diethyl 3,B-benzoyloxy-20-acetoxy-16,2l-hydrazo- 5, 17 (20 -pregnadiene-N ,N -dicanboxylate,

(3') Diethyl 3a,20-diacetoxy 11-keto-16,21 hydrazo- 17 20 -pregnene-N ,N -dicarboxylate,

(4) Diethyl 3 3,20 diacetoxy 11-keto-16,2l-hydrazo- Set-17(20)-pregnene-N ,N -dicarboxylate,

(5) Diethyl 30:,20 diacetoxy 16,21-hydrazo-17(20)- pregnene-N ,N -dicarboxylate,

(6') Diethyl 3,3,20-diacetoxy-16,2l-hydrazo-5a-17(20)- pregnene-N ,N -dicarboxylate,

(7') Diethyl 36,1120 triacetoxy 16-21-hydrazo-5a- 9 l 1 17 (20) -pregnadiene-N ,N -dicarboxylate,

(8) Diethyl 35,11a,20-triacetoxy-16,21-hydrazo 5a- 17 (20)-pregnene-N ,N -dicarboxylate, and

(9) Diethyl 3a,11/3,20-triacetoxy-16,2l-hydrazo 5o:- 17 (20 -pregnene-N ,N -dicarboXylate.

Each of the foregoing 16,21-hydrazo-20-enol acetate derivatives of my invention, upon hydrolysis (such as with methanolic sodium bicarbonate) are converted to the corresponding 20-hydroxy-enol derivative (Le. a compound of Formula V where R is hydrogen, Z being ethoXy) and are isolated in the tautomeric 20-keto form of Formula VI which are named, respectively, as follows:

(1") Diethyl 3,8-acetoXy-20-keto-l6,21-hydrazo 17(a and B)-5-pregnene-N ,N -dicarboxylate,

(2") Diethyl 3B-benzoyloXy-20-keto-16,21 hy'drazo- 17 0c and 13) -5-pregnene-N ,N -dicarboxylate,

(3") Diethyl 3a-acetoxy-11,20-diketo-16,21 hydrazo- 17(a and 5) -pregnene-N ,N -dicarboxylate),

(4") Diethyl 3;8-acetoXy-11,20-diketo-16,21 hydrazo- Soc-17(a and ,8)-pregnane-N ,N -dicarboXylate,

(5") Diethyl 3 3-acetoXy-20-keto-16,21-hydrazo 5a- 17 (oz and B)-pregnane-N ,N -dicarboxylate,

(6") Diethyl 3fi-acetoxy-11,20-diketo-16,21 hydrazo- Soc-17(1): and ,8)-pregnane-N ,N -dicarboxylate,

(7") Diethyl 35,110; diacetoxy-ZO-keto 16,21 hydfaZO-cc-l7(oc and B)-pregnane-N ,N -dicarboxylate, and

(8") Diethyl 3:1,1 1,8-diacetoxy--keto-16,21-hydrazo- 17(a and ,8) -pregnane-N ,N -dicarboxylate.

By way of example, the preparation of a preferred compound of my invention is shown below utilizing 3pbenzoyloxy-S,16-pregnadien-20-one ZO-enol acetate (VII) as starting compound:

Thus, Bfl-benzoyloxy-S,16-pregnadien-20-one 20-en0l acetate (VIII) upon reaction with diethyl azodicarboxylate (II) in benzene for about 6-7 hours according to my process, yields diethyl 3B-benzoyloxy-ZO-acetoxy-16,21- hydrazo-5,17(2O)-pregnadiene-N ,N -dicarboxylate (V") comprising a mixture of the respective 16u,21-hydrazo and 165,21-hydrazo isomers, namely diethyl 3,8-benzoyloxy-20-acetoxy-16a,21-hydrazo 5,17(20) pregnadiene- N ,N -dicarboXylate and diethyl 3,8-benzoyloxy-20-acetoxy-16B,21-hydrazo 5,17(20) pregnadiene N ,N -dicarboxylate, which are separated via chromatographic techniques such as by chromatography over silica gel eluting with benzene-ether.

The enol acetate derivative V" is conveniently converted to the corresponding enol derivative (equivalent to the keto derivative VI) by reaction thereof in refluxing methanolic sodium bicarbonate for about 10 minutes to yield diethyl 3fl-benxoyloxy-20-keto-16,21-hydrazo- 17(oc and p)-5-pregnene-N ,N -dicarboXylate (VI), i.e.

VI'(a, a)

----NG O O CzH and In converting the 20-enol acetate-16,21-hydrazo-17(20)- dehydro adduct derivative (e.g. V") to the 20-keto tautomeric form of the corresponding 20-enol (i.e. to the 20- ket0-16,21-hydrazo-17(a and B) -pregnane derivative, VI) any ester group such as at C3 may or may not be hydrolyzed depending upon the ester group and/or the length of time in which the ester is heated in methanolic sodium bicarbonate. Thus, for example, when diethyl 318, 20 diacetoxy 16,21-hydrazo-5,17(20)-pregnadiene-N N -dicarboxylate in methanolic sodium bicarbonate is heated at reflux temperature for about 15 minutes or less, there is formed diethyl 3 3-acetoxy20-keto-16,2l-hydrazo- 17(a and fi)-5-pregnene-N ,N -dicarboxylate, whereas upon continued heating, the corresponding 3-hydroxy derivative is formed. In general, the 3,8benz0yloxy esters do not hydrolyze as easily as do the corresponding 3-acetoxy esters.

It has been found that the preferred 1,4-adducts of my invention, namely, dilower alkyl 20-alkanoyloxy-l6,21- hydrazo-17(20)-pregnene-N ,N -dicarboxylates as exemplified by diethyl 3,8 benzoyloxy-ZO-acetoxy-16,21-hydraZo-5,l7(20)-pregnadiene-N ,N -dicarboxylate (V) as well as the corresponding 20-hydroxy derivative (isolated in the 20-keto tautomeric form) e.g. diethyl 318-benzoyloxy-20-keto-16,21-hydrazo-17(a and B)-5 pregnene- N ,N -dicarboxylate (VI') and the analogous 3/3-acetoxy derivatives advantageously possess anti-androgenic properties when, for example, they are tested in rats via the subcutaneous route in doses as small as 10 mgm. or 20 mg. per kilogram body weight via pharmacological test procedure similar to that published as Assay No. 17A (effective Apr. 15, 1957) on page 84, Cancer Chemotherapy Reports No. 1, US. Department of Health, Education, and Welfare, Public Health Service. My novel 20- oxygenated 16,21 hydrazopregnane-N ,N -dicarboxylic acid derivatives are thus useful in counteracting the effects of androgen-induced conditions or states and as such may be used in the control and treatment of benign prostatic hypertrophy in dogs. This anti-androgenic activity is exhibited by the isomeric mixtures of the :,21- and 165, 21-hydrazo derivatives exemplified by compounds V and VI above, and is also found to be present in each of the 16mand 16fi-isomers per se. Thus, for example, diethyl 3 3 benzoyloxy-ZO-keto-160:,21-hydrazo-17a-5-pregnene- N ,N dicarboxylate and diethyl 3/3-benzoyloxy-20-keto- 16,8,21 hydrazo 5 pregnene-N ,N -dicarboxylate each alone or when together as an isomeric mixture exhibit anti-androgenic activity in rats via the subcutaneous route at mg-m./kgrn.

The above has been given by way of illustration, it being understood that any -alkanoyloxy-16,20-pregnadiene upon reaction with a dilower alkyl azodicarboxylate or equivalent thereof according to my process will form an isomeric mixture of 20-alkanoyloxy-16,21-hydrazo-17 (20)-pregnene-N ,N -dicarboxylates which are active antiandrogens and which (upon reaction with methanolic sodium bicarbonate) are convertible to the 20-keto-17,20- dihydro tautomeric form of the corresponding 20-hydroxy derivatives which, in turn, are also active as anti-androgens. Specifically, the preferred 16,21-hydrazo-pregnane- N ,N -dicarboxylic acid anti-androgens of my invention include diethyl 3B acetoxy-20-keto-16,21-hydrazo-17(a and 3)-5-pregnene-N ,N -dicarboxylate, diethyl 3,8,20-diacetoxy 11 keto-16,21-hydrazo-5u-17(20) -pregnene-N N xlicarboxylate, diethyl :,20-(113C6t0XY-11-kCtO-16,21 hydrazo-5fi-17(20)-pregnene-N ,N -dicarboxylate, and diethyl 3B-benzoyloxy-20-keto-16,21-hydrazo-17(a and F?)- 5-pregnene-N ,N -dicarboxylate.

My novel 16,21-hydrazo pregnane-N ,N -dicarboxylic acid derivatives as defined by Formulae V and VI can be used in the form of pharmaceutical preparation for subcutaneous administration. In formulating these pharmaceutical compositions, a novel compound of this invention, e.g. diethyl 3B-20-diacetOxy-16,2l-hydrazo-5,l7(20)- pregnadiene-N ,N -dicarboxylate and the 20-hydroxy analog thereof (in the form of the ZO-keto tautomer) i.e. diethyl 3B-acetoxy20-keto 16,21-hydrazo-17(a and B)-5- pregnene-N ,N -dicarboxylate may be used as an isomeric mixture (or a single isomer may be used alone) and is usually compounded with a diluent which is chemically inert to the aforenamed 16,21-hydrazo-N ,N -dicarboxylate.

Preferred as diluents for the aforementioned formulations are aqueous carboxymethylcellulose (containing 0.9% sodium chloride, 0.4% polysorbate 80, 0.5% carboxymethylcellulose and 0.9% benzyl alcohol) and sesame oil (in which is incorporated less than 5% of a solubilizing agent such as dimethylacetamide).

The dosage used of the aforementioned pharmaceutical preparations will vary depending on the nature and severity of the androgen-induced state being treated.

The following examples are representative of my novel process and some of the products produced thereby:

EXAMPLE 1 Condensation of diethyl azodicarboxylate with A -cho lestadiene with formation of diethyl 2a,5a-hydrazo-A cholestene-N ,N -dicarboxylate and diethyl 3B (and 3a) -hydrazino-A -cholestadiene-N ,N -dicarboxylate Reflux a solution of diethyl azodicarboxylate (1.06 g., 0.0051 mole) and A -cholestadiene (2.684 g., 0.0073 mole) in benzene (20 ml.) for 30 hours. The color of the solution gradually fades from orange to pale yellow. Concentrate the solution in vacuo to give a pale yellow oil (wt. 3.30 g.). Chromatograph the oil on neutral alumina (Woelm, grade III) (110 g.), and elute with 1:1 petroleum ether-benzene (300 ml.) obtaining colorless oil (wt. 1.309 g.). Rechromatograph half of the oil on silica gel (66 g.) and elute with benzene (1200 ml.) obtaining diethyl 2a,5a hydrazo-A -cholestene-N ,N -dicarboxylate (351 mg), which is chromatographically homogeneous as determined by thin layer chromatograph,

U.V. kffifigf z transparent above 220 mu Continue the rechromatography of the reaction product with 9:1 benzene-ether (600 ml.) as eluant obtaining diethyl 3,8-hydrazino-A -cholestadiene-N ,N -dicarboxylate as crystals, M.P. 83-92", (121 mg). Recrystallize from 10 aqueous ethanol to obtain colorless needles, M.P. 94-97. Analysis.Calcd. for C H O N C, 73.02; H, 10.03; N, 5.16. Found: C. 73.02; H, 9.83; N, 5.15. [M +31 (c. 0.96 in CHCl U.V. 1322?: transparent above 220 m Elution of the original chromatogram of the reaction product on alumina with 1:1 petroleum ether-benzene (1600 ml. aflords another crop of diethyl 3B-hydrazino- A -cholestadiene-N ,N -dicarboxylate, M.P. 89-93 (182 mg.) which upon recrystallization from aqueous ethanol yields material of M.P. 94-97".

Further elution of the original chromatogram with benzene (1800 ml.) affords diethyl 3a-hydrazino-A -cholestadiene-N ,N -dicarboxylate as crystals, M.P., 147-150", (205 mg). Recrystallization from petroleum ether yields colorless prisms, M.P. 149-151 Aanlysis.-Calcd. for C H O N C, 73.02; H, 10.03; N, 5.16. Found: C, 73-13; H, 10.01; N, 5.25. [M +48 (c. 1.02 in CHCl U.V. x323 transparent above 220 mp.

EXAMPLE 2 Condensation of diethyl azodicarboxylate with -benzoyloxy-n -cholestadiene with formation of diethyl 3 3- benzoyloxy 7 hydrazino-A -cholestadiene-N ,N dicarboxylate and diethyl 3B-benzoyloxy-5u,8a-hydrazo-A -cholestene-N ,N -dicarboxylate Reflux a solution of diethyl azodicarboxylate (500 mg, 0.0029 mole) and 3fi-benzoyloxy-A -cholestadiene (1.30 g., 0.0027 mole) in benzene (26 ml.) for 27 hours. Concentrate in vacuo to obtain a yellow oil (1.80 g.). Chromatograph the oil on silica gel (Davison Co.) (54 g.), and elute with 49:1 benzene-ether (750 ml.) obtaining a colorless oil (486 mg). Rechromatograph the colorless oil on 1 mm. silica gel layer (Kiesegel G nach Stahl, Merck) using benzene as eluant, obtaining diethyl 3B-ben- ZOYlOXY-Su,8oc-hYClIaZO-Sa-A -ChOIESiCIIC N ,N dicarboxylate as colorless needles. M.P. 106-107.5 (72 mg.). Recrystallize from methanol, M.P. 1l0-112 C.

Analysis.-Calcd. for C H O N C, 72.47; H, 8.82; N, 4.23. Found: C. 72.50; H, 8.95; N, 4.37.[1] +44 U.V. M5 3? 230 III 1.

Further elution with 19:1 benzene-ether (790 ml.) affords diethyl 3fl-benzoyloxy 7 hydrazino-A cholestadiene-N ,N -dicarboxylate as colorless needles, M.P. l66-169 C. (628 mg.) which upon recrystallization from methanol yields materials of M.P. 167-170.

Arralysis.-Calcd. for C H O N C, 72.47; H. 8.82; N, 4.23. Found: C, 72.45; H, 8.40; N, 4.25, --38 (c. 0.70 in CHCl U.V. A232? 230 my EXAMPLE 3 Diethyl-3fi-hydroxy-ZO-keto-16,21-hydrazo-17 (a and ,6)-

5-pregnene-N ,N -dicarboxylate and the 3-acetate ester thereof A. CONDENSATION OF DIETHYL AZODICARBOXYLATE WITH 313,20 DIACETOXY 5,16,20 PREGNATRIENE WITH FORMATION OF DIE'IHYL 313,20-DIACETOXY- 16,21 HYDRAZO-5,1T(2O)-PREGNADIENE-N1,N2-DICAR. BOXYLATE To a solution of 36,20-diacetoxy-5,16,20-pregnatriene (700 mg.) in benzene (40 ml.), and 310 mg. of diethyl azodicarboxylate. Reflux the mixture for six and one-half hours, then concentrate in vacuo to give a yellow semisolid residue. Chromatograph this residue on silica gel g.) eluting first with 49:1 benzene-ether (240 ml.),

and then obtaining an unreacted 35,20-diacetoxy-5,16,20- pregnatriene (7 mg). Elute further with 19:1 benzeneether (200 m1.) obtaining a colorless semi-solid (5 mg), and then continue eluting With 19:1 benzene-ether (1180 ml.), obtaining diethyl 35,20-diacetoxy16,2l-hydrazo-S, 17(20)pregnadiene-N N -dicarboxylate (i.e. a mixture comprising diethyl 35,20-diacetoxy 16a,21 hydrazo-5, 17(20)pregnadiene-N ,N -dicarboxylate and diethyl 35, N -dicarboxylate), as a thin layer chromatographic homogenous colorless semi-solid, M.P. 8497 C. (weight 940 mg., 84.8%), [11],; 58" (chloroform):

1%,"; EtOh 199.0 111 1 (e 8860); 1213i? 1754 (sh), 17311711 (S, broad) B. DIETHYL S S-HYDROXYQO KETO 16,21 HYDRAZO- 17(a. AND B)-5-PREGNENE-N1,N2-DICABOXYLATE To a solution of 300 mg. of diethyl 35,20-diacetoxy- 16,21 hydrazo 5, 17(20) pregnadiene-N ,N -dicarboxylate in 90% methanol (50 ml.), add 100 mg. of sodium bicarbonate. Heat the mixture (pH:9.5) at reflux temperature for 40 minutes (at which time the pH of the solution is around 7.5). Add acetic acid dropwise until the pH of the solution is around 6.0. Concentrate in vacuo to a small volume, then extract with ether. Wash the ethereal solution with Water, then dry over anhydrous sodium sulfate, filter and concentrate in vacuo to a pale yellow semisolid residue (weight 268 mg). Chromatograph the residue on silica gel (14 g.). Elution with 9:1 benzene-ether (120 ml.) gives unreacted diethyl 35,20 diacetoxy 16,21 hydrazo 5,16(21) pregnadiene-N ,N -dicarboxylate (16 mg.). Further elution with 7:3 benzene-ether (175 ml.) gives diethyl 35-hydroxy- ZO-keto 16, 21 hydrazo 17(0c and 5) 5 pregnene- N ,N -dicarboxylate (i.e., an isomeric mixture comprising diethyl 35 hydroxy 20 keto 16a,21-hydrazo- 170:,5 pregnene N ,N dicarboxylate and diethyl-35- hydroxy 20 keto 165,21 hydrazo 5 pregnene- N ,N -dicarboxylate), as a thin layer chromatographic homogenous colorless semi-solid, M.P. 5282 C. [a] 40 (chloroform) 11115.? 8465-3395 (m., broad), 1737 (sh), 1727-1695 (3., broad).

C. DIETHYL 3B-ACETOXY20-KETO 16,21 HYDRAZO- 17 (0. AND ,8)-5-PREGNENEN1,N2-DICARBOXYLATE To a solution of 800 mg. of 35 hydroxy- 20 keto- 16,21 hydrazo 17(a and 5) 5 pregnene N ,N dicarboxylate in 12 ml. of pyridine at about C., add acetic anhydride (4 g.). Allow the mixture to stand at room temperature for one hour, then pour into ice water. Extract the aqueous mixture With ether, and wash the ethereal solution first with 10% sulfuric acid followed by water, saturated aqueous sodium bicarbonate solution, and finally again with Water. Dry the washed ethereal solution over anhydrous sodium sulfate, filter and concentrate in vacuo to a solid comprising diethyl 35-acetoxy 20 keto 16,21 hydrazo 17(a and 5- pregnene-N ,N -dicarboxylate (i.e., an isomeric mixture comprising diethyl 35 acetoxy 20 keto 16u,21-hy drazo 170:,5 pregnene N ,N dicarboxylate, and diethyl 35 acetoxy 20 keto 165,21 hydrazor5- pregnene-N ,N -dicarboxylate).

Alternatively, the compound of this example is prepared by treating diethyl 35,20 diacetoxy 16,21 hydrazo 5 pregnene 5,17(20) pregnadiene N ,N dicarboxylate (350 mg.) in 90% methanol (50 ml.). Add a solution of one equivalent weight of sodium bicarbomate in methanol and stir for about ten minutes. Isolate and purify the compound of this example via chromatographic techniques.

EXAMPLE 4 Diethyl 35-benzoyloxy-20-keto-16,21-hydrazo-17 (a and 5)-pregnene-N ,N -dicarboxylate A. CONDENSATION OF DIETHYL AZODICARBOXYLATE 12 WITH 35 BENZOYLOXY-ACETOXY-5,16,20 PREGNA- TRIENE WITH FORMATION OF DIETHYL 3B-BEN- ZOYLOXY-20-AXETOXY 16,21 HYDRAZO 5,17(20) PREGNADIENE-N1,N2-DICARiBOXYLATE Dissolve 5.23 g. of 35-benzoyloxy 20 acetoxy-5,16, 20-pregnatriene and 1.99 g. of diethyl azodicarboxylate in 350 ml. of benzene. Heat the solution at reflux temperature for six hours, then concentrate in vacuo to a residue. Chromatograph the residue on silica gel (236 g.). Elute first with 97:3 benzene-ether (500 ml.) to give 35- benzoyloxy 20 acetoxy 5,16,20 pregnatriene (330 mg). Secondly, elute with 19:1 benzene-ether (2 liters) to give diethyl 35 benzoyloxy 20 acetoxy 16,21- hydrazo 5,17(20) pregnadiene N N dicarboxylate (i.e., a mixture comprising diethyl 35 benzoyloxy-20- acetoxy 1601,21 hydrazo 5,17 (20) pregnadiene- N ,N -dicarboxylate, and diethyl 35 benZoyloxy-20-acetoxy 165,21 hydrazo 5,17(20) pregnadiene N ,N dicarboxylate). (Weight 5.510 g.) M.P. 85.99 C.; 5 1 (chloroform) k'fzzethanol 230 E 16,000), 276 E 2120), 282 m (6 2210);

17371712 (5., broad), 1604 (2), 1575 (w.) cm."

B. DIETHYL -BENZOYLOXY-20-KETO-16,21-HYDRAZO- 17 ((1. AND B) -fi-PREGNENllN NeDlCARBOXYLATE (1) To a solution of diethyl 35 benzoyloxy-ZO-acetoxy 16,21 hydrazo 5,17(20) pregnadiene N N dicarboxylate (3.85 g.) in methanol (400 ml.), add 510 mg. of sodium bicarbonate. Heat the mixture at reflux temperature for 15 minutes, during which time the pH of the solution changes from about 9.5 to about 7.5. Add acetic acid dropwise to the solution until the pH is adjusted to about 6.0, then concentrate the solution in vacuo to a small volume. Add Water and extract with ether. Wash the combined ether extracts with water, then dry over anhydrous sodium sulfate. Concentrate in vacuo to a residue (weight 3.754 g.) comprising diethyl 35- benzoyloxy 20 acetoxy-16,21-hydrazo-5,17(20)-pregnadiene N ,N dicarboxylate (i.e., a mixture comprising diethyl 35 benzoyloxy 20 keto 160:,21 hydrazo- 17a-5- pregnene N ,N dicarboxylate and diethyl 35- benzoyloxy 20 keto 165,21 hydrazo-S-pregnene- N ,N -dicarboxylate) (2) Diethyl 35 benzoyloxy 20-keto-16u,21-hydrazol7ot-5-pregnene N ,N dicarboxylate.-Recrystallize the isomeric mixture obtained in the preceding paragraph (Example 4B-1) from methanol to give diethyl 35-ben' zoyloxy 20 keto 1671,21 hydrazo 17a 5 pregnene- N ,N -dicarboxylate (weight 1.563 g.), M.P. 134 C. Concentrate the filtrate to a residue (weight 2.19 g.) and chromatograph this residue on silica gel (2.19 g.). Elute first with 24:1 benzene-ether (1 liter), and concentrate the eluate to a residue comprising diethyl 35-benzoyloxy- 20 keto 1601,21 hydrazo 17oz 5 pregnene-N ,N dicarboxylate (weight 806 mg), M.P. 113-143" C. The combined yield of diethyl-35-benzoyloxy-ZO-keto-16a,21- hydrazo-17a-5-pregnene-N ,N -dicarboxylate equals 2.369 g. (74.3% theory). Purify by recrystallization from methanol, M.P. 153154 C. [u] 20.7 C. (chloroform) kfizgf 201 (6 27,300), 230 (6 15,900), 274 (e 1170) 280 my 811. pm 1737 5.), 1720 5. 1713 5.), 1604 (w.),

1494 (W.) cm.-

24:1 benzene-ether (1200 ml.) and evaporate the elute to a residue comprising diethyl 3B-benzoyloxy-20-keto- 163,21 hydrazo pregnene N ,N dicarboxylate (Weight 540 mg. 11.5%), M.P. 151163 C. Purify by recrystallization from methanol, M.P. l74l75 C. [1!]; +6 (chloroform) kijfgf 201 (5 28,200), 230 (6 15,200), 274 (e 1170), 280 mu ($11.); 11:3 1720 (s.), 1706 (s.), 1602 (W.), 1586 (W.), 1488 (W.) GILL-1 C. ALTERNATE PROCEDURE FOR THE PREPARATION OF DIETHYL 35-BENZOYLOXY 20 KETO 16,21 HY- DROZO-17(a AND ,8)-5-PREGNENE-N1,N2DICARBOXYL- ATE (1) To a solution of 800 mg. of diethyl-3B-hydroxy-20- keto 16,21 hydrazo 17(0: and 6) 5 pregnene N N -dicarboxylate (prepared as described in Example 313) in 12 ml. of pyridine cooled to about 0 C., add 1.60 g. of benzoyl chloride. Allow the mixture to warm to room temperature and stand at room temperature for one hour. Pour the reaction mixture into ice Water, extract with ether and wash the combined ether extracts With sulfuric acid, Water, saturated aqueous sodium bicarbonate solution, and finally Water. Dry the Washed ether solution over anhydrous sodium sulfate, and concentrate in vacuo to a residue (M.P. 98-107 C., Weight 1.152 mg.) comprising diethyl 3fl-benzoyloxy-20-keto-16,2l-hydrazo-170i and B)-5-pregnene-N ,N -dicarboxylate (i.e. a mixture comprising diethyl3 ,B-benzoyloxy--keto-16a,21- hydrazo-17a-5-pregnene-N ,N dicarboxylate, and diethyl- 36 benzoyloxy 20 keto 165,21 hdrazo 5 pregnene-N ,N -dicarboxylate) Purify by recrystallization from ether to give colorless crystals (M.P. 108-112 C., 701 mg.).

(2) Diethyl 3B benzoyloxy 20 keto 160;,21 hydrazo 17a 5 pregnene N ,N dicarboxylate. Chromatograph the diethyl 3/3-benzoyloxy-20-keto-16,21- hydrazo 17(a and [3) 5 pregnene-N ,N -dicarboxylate (M.P. 108-112 C.), prepared as described in above Example 4C-1, on silica gel (70 g.) eluting with 97:3 benzene-ether (700 ml.). Concentrate the eluate to a residue (M.P. l081ll C., 520 mg.), and recrystallize this residue from methanol to give diethyl-354:enzoyloxy-20- keto 16a,21 hydrazo 17a 5 pregnene N ,N -dicarboxylate, M.P. 141-144 C. (434 mg.). Upon a second recrystallization from methanol, the melting point is 146- 152 C. [111 20.7 (chloroform). The ultra-violet and infrared spectra of this sample are identical to that of the compound prepared as described in Example 4B-2.

(3) Elute the silica gel column prepared in above Example 4C-2 with a second reaction (100 ml.) of a 97:3 benzene ether solvent mixture. Combine the eluates to give a residue for about 33 mg. of diethyl 3/3-benzoyloxy- 20 keto 16,21 hydrazo 17(a and 5) 5 pregnene- N ,N -dicarboxylate.

(4) Diethyl 36 benzoyloxy 20 keto 166,21 hydrazo 5 pregnene N ,N dicarboxylate.-Elute the silica gel column prepared in above Example 4C2 a third time with 350 ml. of 97:3 benzene-ether. Concentrate the eluate to a residue (M.P. 97100 C., 67 mg.). Recrystallize the residue from methanol to give diethyl 3 3 benzoyloxy 20 keto 16 3,21 hydrazo 5 pregnene-N ,N -dicarboxylate (M.P. 147155 C., mg.), upon a second crystallization from methanol, M.P. 173- 174 C. The ultra-violet and infrared spectra of this sample are identical to the compound prepared as described in Example 4B4.

EXAMPLE 5 3fi-hydroxy-20-keto-16,21-hydrazo-17(a and ,8)-5-pregnene-N ,N -dicarbonamide A. CONDENSATION OF AZODICARBONAMIDE WITH 3B,20DIACETOXY-5,16,20-PREGNATRIENE WITH FOR- MATION OF 3B,20DIACETOXY 16,21 HYDRAZO-5,17 (20)-PREGNADIENE-N1,N2-DICARBONAMIDE To a solution of 36,20-diacetoxy-5,16,20-pregnatriene (1.20 g.) in 60 ml. of benzene, add 370 mg. of azodicarbonamide, and heat the mixture at reflux temperature for four hours. Concentrate in vacuo to a residue, add warm chloroform to the residue, and filter. Concentrate the filtrate in vacuo to a residue (1.38 g.) and chromatograpn this residue on silica gel (42 g.), eluting first with 1:1 benzene ether (200 ml.). (Concentration of this eluate yields 3B,20diacetoxy-5,l6,20-pregnatriene, M.P. 136.5- l41.5 C.) Elute the silica gel column a second time with 2:1 methanol (500 ml.). Concentrate the eluate to a residue (M.P. 164-19l C., 334 mg.), and recrystallize this residue from ethanol to give 3/3,20-diacetoxy-16,21- hydrazo 5,17(20) pregnadiene N ,N dicarboxylate (i.e. a mixture comprising 36,20-diacetoxy-l6a,2l-hydrazo 5,17(20) pregnadiene N ,N dicarbonamide, and 35,20 diacetoxy 165,21 hydrazo 5,17(20) pregnadiene-N ,N -dicarbonamide), M.P. 270 C. (decomp.) Weight 180 mg.

kfig 209.5 mu (6 6580); 112$? 3442 (s.), 3326 (s.), 32 24 (sh.), 1755 (s), 1734 (s), 1722 (s.), 1705 (s.), 1684 (s.), 1666 (s.), 1581 (s.), 1558 (sh.) om.

B. 3B-HYDROXY-20KETO-16,21-HYDRAZO-17(a AND ,6)-

5-PREGNENE-N1,Nz-DICARBONAMIDE Reflux for one hour a mixture of 313,20-diacetoxy-16, 2l-hydrazo-5,17(20)-pregnadiene-N ,N dicarbonamide (200 mg.) and sodium bicarbonate (66 mg.) in methanol (66 ml.). Add acetic acid dropwise until the pH of the mixture is adjusted to 7.0, then concentrate the mixture in vacuo to a small volume and extract with chloroform. Wash the combined chloroform extracts With Water, dry over sodium sulfate and concentrate in vacuo to a residue comprising 3,8-hydroxy-20-keto-16,21-hydrazo-1701 and ,8)-5-pregnene-N ,N -dicarbonamide (i.e. a mixture comprising 3B-hydroxy-20-keto-l6a,21-hydrazo- 17a-5-pregnene-N ,N -dicarbonamide, and 3fi-hydroxy- 20-keto-165,21-hydrazo-5pregnene N ,N dicarbonamide). Purify by triturating the residue with Warm chloroform and filtering 00? the resultant precipitate (M.P. 25 0260 (decomp.) followed by recrystallization from methanol (M.P. 256258 (decomp.)), weight 84 mg, and a second recrystallization from methanol (M.P. 258- 259 (decomp.)),

11:35,? 3488 (m.), 3451 (s.), 8270 (s.), 3218 (s.), 3184 (111.) 3146 (111.), 1710 (m.), 1692 (s.), 1682-1860 (s, broad), 1601 (s.), 1583 (s.), 1560 (sh.) cmr EXAMPLE 6 Diethyl 3u-hydroxy-1 1,20-diketo-16,21-hydrazo-pregnane- 11,20-diketo 16,2l-hydrazo-pregnane N ,N -dicarboxylate A. CONDENSATION OF DIETHYL AZODICARBOXYLATE WITH 3a,20-DIACETOXY-11-KETO-16,20-PREGNADIENE WITH FORMATION OF DIETHYL 3a,20-DIACETOXY- 11KETO16,21 HYDRAZO 17(20) PREGNENE N1,N2- DICARBOXYLATE In a manner similar to that described in Example 3A, treat 3a,20-diacetoxy-11-keto-16,20-pregnadiene with diethyl azodicarboxylate in benzene. Isolate and purify the resultant product in a manner similar to that described to give diethyl 30,2O diacetoxy-ll-keto 16,21 hydrazo- 17(20)-pregnene-N ,N -dicarboxylate (i.e. a mixture comprising diethyl 3 a,20-diacetoxy-l 1-keto-16a,21-hydrazo- 17(20)-pregnene-N ,N -dicarboxylate and diethyl 30:,20- diacetoxy-l1-keto-166,21-hydrazo 17(20) pregnene- N ,N -dicarooxy1ate.

B. DIETHYL 3a-I-IYDROXY-11,20DIKETO-16,21-HYDRAZO- 17 (:2. AND B)-PREGNENE-N1,Nz-DICARBOXYLATE In a manner similar to that described in Example 3B, treat diethyl-3a,20-diacetoxy-1l-keto 16,21 hydrazo- 17(20)-pregnene-N ,N -dicarboxylate with sodium bicarbonate in methanol at reflux temperature for 40 minutes. Isolate and purify the resultant product in a manner similar to that described to give 3a-hydroxy-11,20-diketo- 16,2l-hydrazo-l7( x and ,8)-pregnane-N ,N -dicarboxylate (i.e. a mixture comprising 3d-hydI'OXy-11,20-diktO-16OL, 2l-hydrazo-l7a-pregnane-N ,N -dicarboxylate and diethyl 3a-hydroxy-11,20-diketo-16fi,21 hydrazo pregnane-N N -dicarboxylate) EXAMPLE 7 Diethyl 3B-hydroxy-11,ZO-diketo-16,21-hydrazo-5oc-17(u and [3)-pregnaneN ,N -dicarboxylate A. CONDENSATION OF DIETHYL AZODICARBOXYLATE WITH 3fi,20-DIACETOXY-1l-KETO-Sa,16,20-PREGNADI- ENE WITH FORMATION OF DIETHYL 3,8,20-DIACE- TOXY-l1 KETO-16,2l-HYDRAZO-5a-17(20) PREGNENE- N1,N2DICARBOXYLATE In a manner similar to that described in Example 3A, treat 313,20-diacetoxy-1l-keto-Sa-16,20-pregnadiene With diethyl azodicarboxylate in benzene at reflux temperature for 6 /2 hours. Isolate and purify the resultant product in a manner similar to that described to give diethyl 3,3,20-diacetoxy-l1-keto-16,21 hydrazo-5a-17(20)-pregnane-N ,N -dicarboxylate (i.e. a mixture comprising diethyl 3B,20-diacetoxy-11-keto-16ot,21-hydrazo-5a-17(20)- pregnene-N ,N -dicarboxylate, and diethyl-3,8,20-diacetoxy 11 keto 16,,8,21 hydrazo-5a 17(20) pregnene- N ,N -dicarboxylate) B. DIE'IHYL 3B HYDROXY 11,20 DIKETO-16,21-HY- DRAZO 5a 17(a. AND B)-PREGNANE N1,N2 DICAR- BOXYLATE In a manner similar to that described in Example 3B, treat diethyl 3,3,20-diacetoxy-l1-keto-16,21-hydrazo-5a- 17(20)-pregnene-N ,N -dicarboxylate with sodium bicarbonate in methanol at reflux temperature for 40 minutes. Isolate and purify the resultant product in a manner similar to that described to give diethyl 3,8-hydroxy-1 1,20- diketo-16,21-hydrazo-5a-l7 (a and fi)-pregnane-N ,N -dicarboxylate (i.e. a mixture comprising diethyl 3B-hydroxy- 11,20-dik6tO-1606,21-1lYdI3ZO-50L-170L-P16gl131'16 N ,N dicarboxylate, and diethyl -hydroxy-11,20-diketo-16fi,21- hydrazo-5a-pregnane-N ,N -dicarboxylate) EXAMPLE 8 A. Condensation of diethyl azodicarboxylate With 3- acetoxy-l6,20-pregnadienes with formation of diethyl 3-acetoxy-l6,2l-hydrazo 17(20)-pregnene N ,N -dicarboxylates In a manner similar to that described in Example 3A, treat each of the following 3-acetoxy-16,20-pregnadienes with diethyl azodicarboxylate in benzene at reflux temperature for about six hours:

3a-acetoxy-l l-keto-l 6,20-pregnadiene, 35,20-diacetoxy-5a-16,20-pregnadiene,

3 u,20-diacetoxy-16,20-pregnadiene, 35,11,20-triacetoxy-9(11),16,20-pregnatriene, 3,8,11,8,20-triacetoxy-5a-16,20-pregnadiene, and 3 a,11,8-20-triacetoxy-16,20-pregnadiene.

Diethyl 3a,20-diacetoxy-16,21-hydrazo-17(20) pregnene-N ,N -dicarboxylate (i.e. a mixture comprising diethyl 3a,20-diacetoxy-16ot,21-hydrazo-17(20) pregnene- N ,N -dicarboxylate, and diethyl 3u,20'-diacetoxy-16fl,2lhydrazo-1700)-pregnene N ,N -dicarboxy1ate) Diethyl 3 3,11,20-triacetoxy-16,2l-hydrazo SOC-9(11), 17(20)-pregnadiene-N ,N -dicarboxylate (i.e. a mixture comprising diethyl 3,8,11,20-triacetoxy-16u,21 hydrazo- 5oc-9(11),17(2O)-pregnadiene-N ,N dicarboxylate, and diethyl 35,11,20 triacetoxy 16B,21hydraz0-5u-9(1l)- 17 20 -pregnadiene-N ,N -dicarb oxylate Diethyl 3 [3,1 1a,20-triacetoxy-16,2l-hydrazo-5u-17(20) pregnene-N ,N -dicarboxylate (i.e. a mixture comprising diethyl 3,8,11a,20-triacetoxy-16a,21-hydrazo-5a 17(20)- pregnene-N ,N -dicarboxylate, and diethyl 3B,1la,20-triacetoxy-165,21-hydrazo-5u-17(20)-pregnene N ,N dicarboxylate), and

Diethyl 3 (1,1113,20-triacetoxy-16,21-hydrazo-5a-17(20)- pregnene-N ,N -dicarboxylate (i.e. a mixture comprising diethyl 3a,11,B,2O triacetoxy-l6a,21-hydrazo-5a-l7(20)- pregnene-N ,N -dicarboxylate, and diethyl 30:,11fiLZQ-tfiacetoxy 16,8,21 hydrazo-5oc-17(2O)-pregnene-N ,N -dicarboxylate) B. Diethyl 3-hydroxy-20-keto--16,21-hydrazo-17 (a and ,8) pregnane-N ,-N -dicarboxylates In a manner similar to that described in Example 3B, treat each of the 20-acetoxy-16,21-hydrazo-17(20)-pregnene-N ,N -dicarboxylates prepared in preceding Example 8A with sodium bicarbonate and methanol. Isolate the respective resultant products in a manner similar to that described in Example 3B to obtain, respectively:

Diethyl 3u-hydroxy-11,20-diketo-16,21-hydrazo 17(oc and B)-pregnane-N ,N -dicarboxylate (i.e. a mixture comprising diethyl 3oz-hydr0XY-11,2G-dik6t0-16a,21 hydrazo- 17ot pregnane-N ,N -dicarboxylate, and diethyl 3u-hydroxy-l1,20-diketo-16fi,21hydrazo pregnane-N ,N -dicarboxylate),

Diethyl 3B-hydroxy-20-keto-16,21-hydraz0-5a-17(u and ,8) pregnane N ,N -dicarboxylate (i.e. a mixture comprising diethyl 3/8 hydroxy-ZO-keto-16a,21-hydrazo-5a- 17a-pregnane-N ,N dicarboxylate, and diethyl 3/3-hydroxy-ZO-keto-165,2l-hydrazo-Sa-pregnane N ,N -dicarboxylate);

Diethyl Set-hydroxy 20-keto-16,21-hydrazo-17(a and fi)-pregnane N ,N dicarboxylate (i.e. a mixture comprising 3a-hydroxy-20-keto-16a,2l-hydrazo-Hot-pregnanc- N ,N dicarboxylate, and diethyl 3a-hydroxy-20-keto- 165,2 l-hydrazo-pregnane-N N dicarboxylate Diethyl 3 3-hydroxy-11,20-diketo-16,2l-hydrazo 541-17 (0: and B)-pregnane-N ,N -dicarboxylate (i.e. a mixture comprising diethyl 3/3 hydroxy-11,20 diketo ;,21- hydrazo 5w17a-pregnane-N ,N -dicarboxylate, and diethyl 3,B-hydroxy-11,20-diketo-16,8,21-hydrazo 50c pregnane-N ,N -dicarboxylate),

Diethyl 3fl-hydroxy-11a-acetoxy-ZO-keto-16,21-hydrazo- Set-17(u and B)-pregnane-N ,N -dicarboxylate (i.e. a mixture comprising diethyl BB-hydroxy-l1u-acetoxy-20-keto- 16a,2l-hydrazo-5a-17a-pregnan6 N ,N dicarboxylate, and diethyl 3 5 hydroxy-11a-acetoxy-20-keto-165,21-hydrazo-S a-pregnane-N ,N -dicarboxylate), and

Diethyl 3a hydroxy 1lfl-acetoxy-ZO-keto-l6,21-hydrazo-5u-17(a and ,8)-pregnane-N ,N -dicarboxylate (i.e. a mixture comprising diethyl 3a-hydroxy-1lfi-acetoxy-ZO- keto-16a,21-hydrazo-5a-17a-pregnane-N ,N dicarboxylate, and diethyl 3a-hydroxy-11/8-acetoxy-20-keto-16fi,21- hydrazo-S a-pregnane-N ,N -dicarb oxylate) EXAMPLE 9 3B-benzoyloxy-20-keto-16,21-hydrazo-17(a and p) -5-pregnene-N ,N -dicarbonamide A. CONDENSATION OF AZODICARBONAMIDE WITH 33- BENZOYLOXY 2O ACE'IOXY-S-PREGNATRIENE WITH FORMATION OF 35 BENZOYLOXY 2O ACETOXY- 16,21 HYDRAZO 5,17 -20 PREGNADIENE N1,N2-

DICARBONAIIIDE To a solution of 3,8-benzoy1oxy 20-acetoxy-5,16,20- pregnatriene (1.08 g.) in benzene (200 ml.), add azodicarboxylate (1.30 -g.). Stir the reaction mixture at reflux temperature for six hours, then concentrate in vacuo to a residue. Add chloroform (ca. 2.00 ml.) to the residue and filter. Concentrate the filtrate in vacuo, and add benzene to the resulting residue. Filter the resulting crystalline solid which separates comprising 3p-benzoyloxy-20-acetoxy-l6,21-hydrazo-5,17(20)-pregnadiene N ,N dicarbonamide (i.e. a mixture comprising 3/3-benzoyloxy-20- acetoxy-16a,21-hydrazo-5,17(20) pregnadiene N ,N dicarbonamide, and 3,8-benzoyloxy-20-acetoxy-l6fl,2l-hydram-5,17(20)-pregnadiene-N ,N -dicarbonamide), M.P. 244-245" C. (decrnp.) Weight 186 mg. (53% yield). Purify by recrystallization from methanol, M.P. 249-253 C. (decomp) 04 -82 (c. 0.53 in chloroform) Uv m IR 11231? cm.-

3464 8. 3417 (8. 3325 (8.), 3299 (8. (NH), 1756 (8. zo-ococn 1728 (811.), 1711-1671 (8. 1580 8. 1562 8. 1554 8. (3c-OC0C H CONHZ), 1604 (w.), 1531 111. 1493 (w.) 0:0

NMR 7' in pyridine: 5.25 (singlet, four protons) (NH),

7.87 (singlet, three protons) (-OCOCH 9.00 (singlet,

three protons) and 9.02 (singlet, three protons) (18- and B. 3B-BENZOYLOXY 20 KETO 16,21 HYDRAZO-17- (0. AND B)-5-PREGNENE-N1,N2-DICARBONAMIDE To a solution of 3B-benzoy1oxy-20-acetoxy- 16,21 hydrazo-S,17(20)-pregnadiene-N ,N dicarbonamide (500 mg.) in 90 ml. of methanol (250 ml.), add 73 mg. of sodium bicarbonate (one equivalent). Reflux the solution for ten minutes (during which time the pH of the solution changes from 8.0 to about 7.5). Cool and add glacial acetic acid dropwise (about eight drops) until the pH is adjusted to about 6.0. Concentrate the solution in vacuo and add water (100 ml.) to the residue. Filter, wash with water, and dry the resultant crystalline product which separates comprising -benzoyloxy-20-keto-1 6,21 hydrazo- 17(a and 9)-5-pregnenc-N ,N -dicarbonamide (i.e. a mixture comprising 33 benzoyloxy-20-keto-16a,2 l-hydrazo- 176t-5-pregnene N ,N -dicarbonamide, and 3 8-benzoy1- oxy-20-keto-16,8,21-hydrazo-5-pregnene-N ,N dicarbonamide), M.P. 242-245" C. (decomp.) weight 439 mg. Purify by recrystallization from 99% ethanol, M.P. 244- 246 C. (decomp.) (highly hygroscopic) weight 399 mg. (86.2%

UV tra IR A231? cm.-

3510-3180 (s.) (NH), 1720-1636 (s.), 1602-1573 (s.)

(3B-OCOC H CONH C 0 0), 1540 (W.), 1520 (w.), 1493 (w.) (C= .t,)

NMR 1- in pyridine: 5.00 (singlet, four protons) (NH), 9.03 (singlet, three protons) and 9.18 (singlet, three protons) (18- and 19-CH I claim:

1. The process which comprises reacting an azodicarboxylic acid lower alkyl ester with a steroid of the pregnane series having a conjugated diene present at position (16,20) and having no other double bond conjugated to said diene, in an inert solvent at a temperature in the range of from about room temperature to the refluxing temperature of the reaction mixture, whereby is formed :1 16,21 hydrazo 17(20) pregnene N ,N dicarboxylic acid lower alkyl ester.

2. The process according to claim 1 wherein said azodicarboxylic acid lower alkyl ester is diethyl azodicarboxylate and wherein said steroid of the pregnane series is a 20-lower alkanoyloxy-16,20-pregnadiene, and there is formed a 20-lower alkanoyloxy-16,21-hydrazo- 17'(20)-pregnene-N ,N -dicarboxylic acid ethyl ester.

3. The process according to claim 2 wherein diethyl azodicarboxylate is reacted with 3fl,20diacetoxy-5,16,20- pregnatriene in refluxing benzene whereby is formed diethyl 3[3,20 diacetoxy 16,21 hydrazo 5,17(20)- pregnadiene-N ,N -dicarboxylate.

4, The process according to claim 2 wherein diethyl azodicarboxylate is reacted with 3 3-benzoyloxy-20-acetoxy-S,16,20-pregnatriene in refluxing benzene whereby is formed diethyl 3,8-benzoyloxy-16,2l-hydrazo-5,l7(20)- prgnadiene-N ,N -dicarboxylate.

5. The process according to claim 2 including the additional step of reacting said 20-lower alkanoxyloxy-16,2lhydrazo-17(20)-pregnene-N N -dicarboxylic acid ethyl ester thereby formed with methanolic sodium bicarbonate whereby is formed a 20-keto-16,20-hydraZ0-17(a and )3)- pregnane-N ,N dicarboxylic acid ethyl ester.

6, The process according to claim 5 wherein diethyl azodicarboxylate is reacted with 3fi-benzoyloxy-20-acet0xy-5,16,20-pregnatriene in refluxing benzene and the diethyl 3,6 'benzoyloxy 20 acetoxy 16,21 hydrazo- 5,17(20) pregnadiene N ,N dicarboxylate thereby formed is heated in methanolic sodium bicarbonate whereby is formed 3/5-benzoyloxy-20-keto-16,21-hydrazo-17- (oz and B)-5-pregnene-N ,N -dicarboxylate.

7. The process according to claim 5 wherein diethyl azodicarboxylate is reacted with 3 3,20-diacetoxy-5,16,20- pregnatriene in refluxing benzene and the diethyl-3-fi,20- diacetoxy 16,21 hydrazo 5,17(20) pregnadiene- N ,N -dicarboxylate thereby formed is treated with sodiumbicarbonate in refluxing methanol whereby is formed diethyl 3,8 hydroxy 20 keto 16,21 hydrazo 17 (a and [5) -5-pregnene-N ,N -dicarboxylate.

8. A compound selected from the group consisting of 16,2l-hydrazo-l7(20)-pregnenes of the following Formula I, the S-dehydro analogs thereof, and when R is hydrogen, the 20-keto-17,20-dihydro tautomeric form thereof of Formula II E I fi N-COZ UNITED STATES PATENT OFFICE CERTIFICATE OF CORRECTION Patent No. 3,438,974 April 15, 19c

Munemitsu Tomoeda It is certified that error appears in the above identified patent and that said Letters Patent are hereby corrected as shown below:

Column 1, lines 28 to 37, that portion of Formula II reading Column 3, line 7, "ecyl ester" 'should read acyl ester Column 4 line 43, "especialy" should read especially Column 6, line 21 (1 Dieth 38,20-diacetoxy-l6,21-hydrazo-5 ,7 (20)pregnadiene-Nl,N2-dicarboxylate" should read (l) 3B-acetoxy-5,16-pregnadien-20-one 20-eno1 acetate 36,20- diaceti 16, 2O- pregnatriene Column 7, lines 36 to 42, "Nal, HCO CH OH" should read NaJ HCO /CH OH line 57, "acetate (VIII) should read acetate (VIIL lin 74, BB-benxoyloxy" should read 3B-benzoyloxy Column 9, line 27 "pharmaceutical preparation" should read pharmaceutical pre arations Column 10, lines 44 and 45 [oz] D 44" should read [d] 7 44 (c 0. 70

in CHCl line 55, "(C. 0.70 in CHCl should read (c 1.00 in CHCl line 71, "and 310 mg. should read Add 310 mg. Column 11, lines 8 and 9, "3B,N -dicarboxylate" should read 38,20-diacetoxy-l66 ,2l-hydrazo-5 l7 (20)pregnadiene-N ,N2--dicarboxylate Column 12, line 3, "20axetoxy" should read 20-acetox'y Column 13, lines 11 and 12, "16,2l-hydrozo" should read 16,2l-hydrazo line 31, "16B, 2lhdrazo" should read l6B,2l-hydrazo Column 16, line 74, acetoxy5 pregnatriene" should read acetoxy5,l6 ,20-pregnatriene Column 17 line 56, nujol max cm should read IR 7323 -l line 60, (C u) should read (C C) Column 18, lines 66 and 67 "consisting hydrogen" should read consisting 0 hydrogen Signed and sealed this 14th day of April 1970.

(SEAL) Attest:

EDWARD M.FLE'ICHER,JR. WILLIAM E. SCHUYLER, JR Attesting Officer Commissioner of Patents 

